The molecular relationships of the IncPl plasmids RPl, RP4, R68 and RK2 were tested by electron microscopic examination of heteroduplexes. In several hybridization experiments molecules were detected which had a 7.8% portion of incomplete reannealing. This ' heterologous region ' could be explained by the typical renaturation behaviour of the transposon Tnl. The identity of the Tnl transposon present in RP1 and RP4 was proved by heteroduplex experiments with h phage DNA containing this transposon. These results indicated that the plasmids RP1 and RP4 are identical. Additional heteroduplex experiments between plasmids R68.45 and RP8 and between R68.45 and RK2 were performed. R68.45, a derivative of R68, has a small DNA insertion and RP8 can be regarded as a large insertion mutant of RP4 ; both insertions were used as single-stranded hybridization markers. From the hybrid molecules formed, it was deduced that R68 and RK2 are identical with RPl and RP4 as far as molecular structure is revealed by the technique used.

No abstract

No abstract

No abstract

SUMMARYPlasmids R68 and R68.45 were transferred fromPseudomonas aeruginosatoEscherichia coliby conjugation. R68.45 was able to mobilize theE. colichromosome from different origins at a frequency of about lO−6/donor cell. With R68, no transfer of chromosomal genes could be detected. Plasmid R68.45 differs from its parent R68 only by an additional DNA segment, 2120 bp long, located close to the kanamycin resistance gene. By restriction enzyme analysis it was shown that the additional DNA segment of R68.45 is a duplication of a pre-existing DNA region of R68. The duplicated region is characterized by the following sequence of restriction sites: A–310 bp–SmaI–70 bp–PstI–795 bp–PstI–15 bp–KpnI–540 bp–HpaI–370 bp–B.The endpoints A and B of the duplicated region were determined by a heteroduplex experiment betweenHindIII linearized molecules of R68 and R68.45. It is proposed that this duplication found in R68.45 is responsible for its chromosome mobilizing ability.